Role In Epigenetic Control Research Articles

The stem cell zinc finger 1 (SZF1)/ZNF589 protein has a human-specific evolutionary nucleotide DNA change and acts as a regulator of cell viability in the hematopoietic system

The stem cell zinc finger 1 (SZF1)/ZNF589 protein belongs to the large family of Krüppel-associated box domain-zinc finger (KRAB-ZNF) transcription factors, which are present only in higher vertebrates and epigenetically repress transcription by recruiting chromatin-modifying complexes to the promoter regions of their respective target genes. Although the distinct biological functions of most KRAB-ZNF proteins remain unknown, recent publications indicate their implication in fundamental processes, such as cell proliferation, apoptosis, differentiation, development, and tumorigenesis. SZF1/ZNF589 was first identified as a gene with SZF1-1 isoform specifically expressed in CD34(+) hematopoietic cells, strongly suggesting a role in epigenetic control of gene expression in hematopoietic stem/progenitor cells (HSPCs). However, the function of SZF1/ZNF589 in hematopoiesis has not yet been elucidated. Our study reveals SZF1/ZNF589 as a gene with a human-specific nucleotide DNA-change, conferring potential species-specific functional properties. Through shRNA-mediated loss-of-function experiments, we found that changes in expression of fundamental apoptosis-controlling genes are induced on SZF1/ZNF589 knockdown, resulting in inhibited growth of hematopoietic cell lines and decreased progenitor potential of primary human bone marrow CD34(+) cells. Moreover, we found that the SZF1/ZNF589 gene is differentially regulated during hypoxia in CD34(+) HSPCs in a cytokine-dependent manner, implicating its possible involvement in the maintenance of the hypoxic physiologic status of hematopoietic stem cells. Our results establish the role of SZF1/ZNF589 as a new functional regulator of the hematopoietic system.

Tex10 Coordinates Epigenetic Control of Super-Enhancer Activity in Pluripotency and Reprogramming

Super-enhancers (SEs) are large clusters of transcriptional enhancers that are co-occupied by multiple lineage-specific transcription factors driving expression of genes that define cell identity. In embryonic stem cells (ESCs), SEs are highly enriched for the core pluripotency factors Oct4, Sox2, and Nanog. In this study, we sought to dissect the molecular control mechanism of SE activity in pluripotency and reprogramming. Starting from a protein interaction network surrounding Sox2, we identified Tex10 as a key pluripotency factor that plays a functionally significant role in ESC self-renewal, early embryo development, and reprogramming. Tex10 is enriched at SEs in a Sox2-dependent manner and coordinates histone acetylation and DNA demethylation at SEs. Tex10 activity is also important for pluripotency and reprogramming in human cells. Our study therefore highlights Tex10 as a core component of the pluripotency network and sheds light on its role in epigenetic control of SE activity for cell fate determination.

Analysis of genetic and epigenetic effects of maize seeds in response to heavy metal (Zn) stress.

Conditions of environmental stress are known to lead genetic and epigenetic variability in plants. DNA methylation is one of the important epigenetic mechanisms and plays a critical role in epigenetic control of gene expression. Thus, the aim of the study was to investigate the alteration of genome methylation induced by zinc stress by using coupled restriction enzyme digestion-random amplification (CRED-RA) technique in maize (Zea mays L.) seedlings. In addition, to determine the effect of zinc on mitotic activity and phytohormone level, high-pressure liquid chromatography (HPLC) and mitotic index analysis were utilized. According to the results, mitotic index decreased in all concentrations of zinc except for 5mM dose and chromosome aberrations such as c-mitosis, stickiness, and anaphase bridges were determined. It was also observed that increasing concentrations of zinc caused an increase in methylation patterns and decrease in gibberellic acid (GA), zeatin (ZA), and indole acetic acid (IAA) levels in contrast to abscisic acid (ABA) level. Especially increasing of ABA levels under zinc stress may be a part of the defense system against heavy metal accumulation in plants.

Hydroxymethylation and DNA methylation profiles in the prefrontal cortex of the non-human primate rhesus macaque and the impact of maternal deprivation on hydroxymethylation

5-Hydroxymethylcytosine (5hmC) is abundant in the brain, suggesting an important role in epigenetic control of neuronal functions. In this paper, we show that 5hmC and 5-methylcytosine (5mC) levels are coordinately distributed in gene promoters of the rhesus macaque prefrontal cortex. Although promoter hydroxymethylation and methylation are overall negatively correlated with expression, a subset of highly expressed genes involved in specific cerebral functions is associated with high levels of 5mC and 5hmC. These relationships were also observed in the mouse cortex. Furthermore, we found that early-life maternal deprivation is associated, in the adult monkey cortex, with DNA hydroxymethylation changes of promoters of genes related to neurological functions and psychological disorders. These results reveal that early social adversity triggers variations in brain DNA hydroxymethylation that could be detected in adulthood.

One bacterial effector with two distinct catalytic activities by different strains

The causative agent of Legionnaires' disease, Legionella pneumophila , is a master manipulator of various eukaryotic hosts ranging from unicellular amoebae to mammals [1]. This intracellular pathogen, which invades macrophages in mammalian hosts, evades the default endosome–lysosome pathway and remodels the phagosome, enclosing it in an endoplasmic reticulum‐derived Legionella ‐containing vacuole (LCV). L. pneumophila facilitates this by hijacking a myriad of eukaryotic cellular functions through translocation of around 300 effectors into the host cell by the Dot/Icm type IVB secretion apparatus. More than 70 of the injected effector proteins contain eukaryotic‐like domains, including the ankyrin repeat, Sel1, F‐box, SET, U‐box and leucine‐rich repeats, which suggests L. pneumophila hijacks and manipulates various eukaryotic processes through molecular mimicry. Elegant work by Rolando et al [2] and Li et al [3] described a new effector, designated RomA or LegAS4, respectively, which exhibits molecular mimicry of host SET‐domain‐containing histone methyltransferases to hijack host transcriptional programming by methylating histones on specific lysine residues [2,3]. Histone methylation has a crucial role in epigenetic control of gene transcription. Prior to these findings, no intracellular pathogen had been shown to directly affect chromatin architecture to alter the host cell transcriptional landscape. Rolando and Li both show that RomA/LegAS4 is a histone methyltransferase, but that the functional phenotype of RomA/LegAS4 is different in the two strains of L. pneumophila used in the two studies. Rolando et al primarily used the …

Acute β-Adrenergic Activation Triggers Nuclear Import of Histone Deacetylase 5 and Delays Gq-induced Transcriptional Activation

During hemodynamic stress, catecholamines and neurohumoral stimuli may induce co-activation of G(q)-coupled receptors and β-adrenergic receptors (β-AR), leading to cardiac remodeling. Dynamic regulation of histone deacetylase 5 (HDAC5), a transcriptional repressor, is crucial during stress signaling due to its role in epigenetic control of fetal gene markers. Little is known about its regulation during acute and chronic β-AR stimulation and its cross-interaction with G(q) signaling in adult cardiac myocytes. Here, we evaluate the potential cross-talk between G(q)-driven and β-AR mediated signaling at the level of nucleocytoplasmic shuttling of HDAC5. We show the translocation of GFP-tagged wild type HDAC5 or mutants (S279A and S279D) in response to β-AR or G(q) agonists. Isoproterenol (ISO) or PKA activation results in strong nuclear accumulation of HDAC5 in contrast to nuclear export driven by Ca(2+)-calmodulin protein kinase II and protein kinase D. Moreover, nuclear accumulation of HDAC5 under acute ISO/PKA signaling is dependent on phosphorylation of Ser-279 and can block subsequent G(q)-mediated nuclear HDAC5 export. Intriguingly, the attenuation of G(q)-induced export is abolished after chronic PKA activation, yet nuclear HDAC5 remains elevated. Last, the effect of chronic β-AR signaling on HDAC5 translocation was examined in adult myocytes from a rabbit model of heart failure, where ISO-induced nuclear import is ablated, but G(q)-agonist mediated export is preserved. Acute β-AR/PKA activation protects against hypertrophic signaling by delaying G(q)-mediated transcriptional activation. This serves as a key physiological control switch before allowing genetic reprogramming via HDAC5 nuclear export during more severe stress, such as heart failure.

Distinct mode of methylated lysine-4 of histone H3 recognition by tandem tudor-like domains of Spindlin1

Recognition of methylated histone tail lysine residues by tudor domains plays important roles in epigenetic control of gene expression and DNA damage response. Previous studies revealed the binding of methyllysine in a cage of aromatic residues, but the molecular mechanism by which the sequence specificity for surrounding histone tail residues is achieved remains poorly understood. In the crystal structure of a trimethylated histone H3 lysine 4 (H3K4) peptide bound to the tudor-like domains of Spindlin1 presented here, an atypical mode of methyllysine recognition by an aromatic pocket of Spindlin1 is observed. Furthermore, the histone sequence is recognized in a distinct manner involving the amino terminus and a pair of arginine residues of histone H3, and disruption of the binding impaired stimulation of pre-RNA expression by Spindlin1. Our analysis demonstrates considerable diversities of methyllysine recognition and sequence-specific binding of histone tails by tudor domains, and the revelation furthers the understanding of tudor domain proteins in deciphering epigenetic marks on histone tails.

A Perspective on Nutrition and Cancer

A Perspective on Nutrition and Cancer Research into nutrition and cancer typically involves either a study of dietary factors in the context of cancer prevention, or dietary changes to moderate the effects of conventional cancer therapy. Cancer involves genetic and epigenetic changes that influence gene expression programs controlling cell growth, death, differentiation. Some of the changes leading to genomic instability can be influenced by diet and other environmental factors. In this context, a genoprotective diet may be considered as one that is low in harmful dietary factors and sufficient in protective dietary factors.Harmful factors, e.g., carcinogens, are ones that can promote cancer. Possible beneficial factors include some vitamins, minerals, and phytochemicals. The B-vitamin folate is an example of a possible protective factor; among other functions, it is involved in dTMP (DNA) biosynthesis and has a role in epigenetic control.

Epigenetic regulation of human embryonic stem cells.

Recently, there has been tremendous progress in characterizing the transcriptional network regulating human embryonic stem cells (hESCs; MacArthur etal., 2009; Loh etal., 2011), including those signaling events mediated by Oct4, Nanog, and Sox2. There is growing interest in the epigenetic machinery involved in hESC self-renewal and differentiation. In general, epigenetic regulation includes chromatin reorganization, DNA modification, and histone modification, which are not directly related to alterations in DNA sequences. Various protein complexes, including Polycomb, trithorax, nucleosome remodeling deacetylase, SWI/SNF, and Oct4, have been shown to play critical roles in epigenetic control of hESC physiology. Hence, we will formally review recent advances in unraveling the multifaceted role of epigenetic regulation in hESC self-renewal and induced differentiation, particularly with respect to chromatin remodeling and DNA methylation events. Elucidating the molecular mechanisms underlying the maintenance/differentiation of hESCs and reprogramming of somatic cells will greatly strengthen our capacity to generate various types of cells to treat human diseases.

Histone deacetylase HD2 interacts with ERF1 and is involved in longan fruit senescence.

Histone deacetylation plays an important role in epigenetic control of gene expression. HD2 is a plant-specific histone deacetylase that is able to mediate transcriptional repression in many biological processes. To investigate the epigenetic and transcriptional mechanisms of longan fruit senescence, one histone deacetylase 2-like gene, DlHD2, and two ethylene-responsive factor-like genes, DlERF1 and DlERF2, were cloned and characterized from longan fruit. Expression of these genes was examined during fruit senescence under different storage conditions. The accumulation of DlHD2 reached a peak at 2 d and 30 d in the fruit stored at 25 °C (room temperature) and 4 °C (low temperature), respectively, or 6 h after the fruit was transferred from 4 °C to 25 °C, when fruit senescence was initiated. However, the DlERF1 transcript accumulated mostly at the later stage of fruit senescence, reaching a peak at 5 d and 35 d in the fruit stored at 25 °C and 4 °C, respectively, or 36 h after the fruit was transferred from low temperature to room temperature. Moreover, application of nitric oxide (NO) delayed fruit senescence, enhanced the expression of DlHD2, but suppressed the expression of DlERF1 and DlERF2. These results indicated a possible interaction between DlHD2 and DlERFs in regulating longan fruit senescence, and the direct interaction between DlHD2 and DlERF1 was confirmed by yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays. Taken together, the results suggested that DlHD2 may act with DlERF1 to regulate gene expression involved in longan fruit senescence.

Willow volatiles influence growth, development, and secondary metabolism in Aspergillus parasiticus

Aflatoxin is a mycotoxin and the most potent naturally occurring carcinogen in many animals. Aflatoxin contamination of food and feed crops causes a significant global burden on human and animal health. However, available methods to eliminate aflatoxin from food and feed are not fully effective. Our goal is to discover novel, efficient, and practical methods to control aflatoxin contamination in crops during storage. In the present study, we tested the effect of volatiles produced by willow (Salix acutifolia and Salix babylonica) and maple (Acer saccharinum) bark on fungal growth, development, and aflatoxin production by the fungus Aspergillus parasiticus, one economically important aflatoxin producer. S. acutifolia bark volatiles nearly eliminated aflatoxin accumulation (>90% reduction) by A. parasiticus grown on a minimal agar medium. The decrease in aflatoxin accumulation correlated with a twofold reduction in ver-1 (encodes a middle aflatoxin pathway enzyme) transcript level. Expression data also indicate that one histone H4 acetyltransferase, MYST3, may play a role in epigenetic control of aflatoxin gene transcription in response to volatile exposure. Volatiles derived from wood bark samples also increased fungal growth up to 20% and/or enhanced conidiospore development. Solid-phase microextraction-gas chromatographic-mass spectrometric analysis of bark samples identified sets of shared and unique volatile compounds that may mediate the observed regulatory effects on growth, development, and aflatoxin synthesis. This work provides an experimental basis for the use of willow industry by-products to control aflatoxin contamination in food and feed crops.

Modulating the Modulator: Epigenetic Control Beyond the Histone Code

Site‐specific post‐translational modifications of nucleosomal histones associated with chromatin structural change have been established to play an important role in epigenetic control of gene transcriptional activation or silencing. Growing evidence from recent studies argues that epigenetic gene transcription is governed not only by modifications of histones and DNA, but also by non‐coding RNAs in the cell nucleus. However, the molecular mechanistic understanding of the latter remains elusive. In this talk, I will present our structural and mechanistic analysis of the functional role of non‐coding RNA transcripts in histone H3 lysine 27 methylation mediated gene transcriptional silencing by Polycomb repressive complexes. I will discuss functional implications of the fundamental epigenetic interplay between RNA and histone lysine methylation in gene repression in human biology and disease.

Isolation and comparative expression analysis of six MBD genes in wheat

The 5-methylcytosines (m5C) play critical roles in epigenetic control, often being recognized by proteins containing an MBD. In this study, we isolated six wheat cDNAs with open reading frame encoding putative methyl-binding domain proteins, which were designated as TaMBD1-TaMBD6, respectively. BLASTX searches and phylogenetic analysis suggested that the six TaMBD genes belonged to four (I, II, III and VIII) of the eight subclasses of MBD family. Genomic analysis showed that a 1386 bp intron was included in TaMBD1 and a 12-bp intron was found in TaMBD4. The expression profiles of the six TaMBDs were studied via Q-RT-PCR and the results indicated that the TaMBDs were differentially expressed in detected wheat tissues. It was interesting to note that 3 TaMBDs were highly expressed in dry seeds and endosperms. Moreover, the differential expression patterns of TaMBDs were observed in leaves and roots under water-stress. We concluded that multiple wheat MBD genes were present and they might play important roles in wheat growth and development, as well as in the water-stress response.

Symptoms of Allergic Rhinitis in Mothers during Early Pregnancy are Associated with Higher Prevalence of Allergic Rhinitis in their Offspring

RATIONALE: Epigenetic control of gene expression profiles is a ubiquitous mechanism during cell differentiation, organogenesis or chronic inflammatory reactions. Recent studies showed that allergen exposure during pregnancy increases bronchial hypersensitivity of their offspring in a murine model of bronchial asthma, however, no such phenomena were reported in humans. In the present study, the role of epigenetic control in the onset of allergic diseases was investigated.

Symptoms of Allergic Rhinitis in Women during Early Pregnancy Are Associated with Higher Prevalence of Allergic Rhinitis in Their Offspring

Epigenetic control of gene expression profiles is a ubiquitous mechanism during cell differentiation, organogenesis and chronic inflammatory reactions. Recent studies have shown that allergen exposure during very early pregnancy increases bronchial hypersensitivity in offspring in a murine model of bronchial asthma. However, no such phenomena were reported in humans. In the present study, the role of epigenetic control in the onset of allergic diseases was investigated. A total of 400 pairs of mothers with physician-diagnosed allergic rhinitis (AR) and their offspring (age 7-18 months) who participated in a large-scale medical check-up were enrolled in this retrospective cohort study. Family history of allergic diseases and the presence or absence of AR symptoms during pregnancy were inquired about using a self-answered questionnaire. A logistic regression model adjusted for age, gender, birth month and father's history of allergic diseases was statistically analyzed. Offspring whose mothers had any AR symptoms during early pregnancy showed a significantly higher adjusted odds ratio for the onset of AR in offspring than those whose mothers had no symptoms during pregnancy (adjusted Odds Ratio: 6.26, p = 0.036). However, the symptoms of AR during late pregnancy showed no effects on the odds ratio. In contrast, the presence or absence of AR symptoms during early or late pregnancy showed no association with the prevalence of food allergy, atopic dermatitis or asthma in offspring. Our results suggest the presence of possible epigenetic mechanisms regulating the onset of AR in humans presumably through increased organ-specific hypersensitivity.

Interaction Between Methyl CpG-Binding Protein and Ran GTPase during Cell Division in Tobacco Cultured Cells

Methyl CpG-binding proteins are considered to play critical roles in epigenetic control of gene expression by recognizing and interacting with 5-methylcytosine (m(5)C) in eukaryotes. However, among 13 corresponding genes in Arabidopsis thaliana, designated as featuring a methyl-binding domain (MBD), only four have so far been shown actually to bind to m(5)C. One example, AtMBD5, was selected here to screen for interacting proteins. Yeast two-hybrid assays were used for screening, and physical interaction was confirmed by pull-down and bimolecular fluorescence complementation (BiFC) assays. Cellular localization was analysed by fluorescence-tagged fusion proteins using tobacco (Nicotiana tabacum) cultured bright yellow 2 cells. A gene finally identified was found to encode AtRAN3, a protein that belongs to the Ran GTPase family, which plays a critical role in nucleocytoplasmic transport and spindle bipolarization during cell division. AtMBD5 and AtRAN3 were clearly shown to interact in the nucleus by BiFC. On co-expression of AtMBD5-cyan fluorescence protein and yellow fluorescence protein-AtRAN3 in tobacco cells, both localized to the nucleus in the resting stage, migrating to the cytoplasm, primarily around chromatin, during mitosis, particularly at metaphase. These results suggest that AtMBD5 becomes localized to the vicinity of chromosomes with the aid of AtRAN3 during cell division, and may play an important role not only in maintenance of chromatin structures by binding to m(5)C, but also in progress through mitosis by detaching from m(5)C. The present findings also shed light on the physiological function of Ran GTPases, direct target proteins of which have not thus far been well defined, suggesting their key role in chromatin movements in plant cells.

Methylated DNA-binding proteins from Arabidopsis.

The 5-methylcytosines (m5C) play a critical role in epigenetic control, often being recognized by proteins containing a methyl-CpG-binding domain (MBD). Database screening has identified at least 12 putative methyl-CpG-binding proteins from Arabidopsis; we have isolated corresponding cDNAs for seven of them. Despite variation in size and amino acid sequence, all seven proteins exclusively migrate into the nucleus as revealed by green fluorescent protein fusion protein assay, suggesting a relationship with chromatin structure. However, DNA-binding assays using bacterially expressed proteins and synthetic oligonucleotides containing m5C in CpGs showed only one to specifically bind, designated AtMBD5. Further analysis showed that AtMBD5 efficiently binds to m5C in CpNpN (N is A, T, or C) but not in CpNpG sequences, both frequently found in plant DNA. The other six proteins showed either nonspecific DNA binding or no ability to recognize m5C. RNA-blot hybridization and immunoblot analysis indicated AtMBD5 to be present essentially in all tissues. Using green fluorescent protein driven by the authentic promoter, AtMBD5 was found to be actively expressed in pistils and root tips. Because m5Cs in CpG and CpNpN are considered to function in gene expression and gene silencing, respectively, the present results suggest that AtMBD5 may have distinct functions in regulation and/or self defense of genes in actively proliferating cells.

Four-dimensional imaging of chromatin dynamics during the assembly of the interphase nucleus.

Large-scale chromatin organization is likely to play an important role in epigenetic control of gene expression. This implies that after mitosis the correct chromatin organization must be re-established in the nuclei of the two daughter cells. Here we analyze the dynamic behavior of chromatin during the transition from late anaphase to G1 in dividing HeLa cells, which express green fluorescent protein-tagged histone H2B. Time-lapse confocal microscopy was used to image the movement and the decondensation of chromatin as cell division progresses. Typically, time series of over 100 three-dimensional images (4D images) were collected, spanning a time period of up to three hours. Special care was taken to avoid photodamage, since cell cycle progression is exquisitely sensitive to photochemical damage. Quantitative analysis of the 4D images revealed that during the anaphase to G1 transition the movement of chromatin domains relative to other chromatin is remarkably limited. Chromatin dynamics can best be described as a radial expansion of the cluster of chromosomes that is present in late anaphase. We find that decondensation occurs in two phases. First a rapid decondensation by about a factor of two, followed by a slower phase in which part of the chromatin does not decondense any further, whereas the remaining chromatin decondenses further about two fold.

Drosophila Chromosome Condensation Proteins Topoisomerase II and Barren Colocalize with Polycomb and Maintain Fab-7 PRE Silencing

Mechanisms of cellular memory control the maintenance of cellular identity at the level of chromatin structure. We have investigated whether the converse is true; namely, if functions responsible for maintenance of chromosome structure play a role in epigenetic control of gene expression. We show that Topoisomerase II (TOPOII) and Barren (BARR) interact in vivo with Polycomb group (PcG) target sequences in the bithorax complex of Drosophila, including Polycomb response elements. In addition, we find that the PcG protein Polyhomeotic (PH) interacts physically with TOPOII and BARR and that BARR is required for Fab-7-regulated homeotic gene expression. Conversely, we find defects in chromosome segregation associated with ph mutations. We propose that chromatin condensation proteins are involved in mechanisms acting in interphase that regulate chromosome domain topology and are essential for the maintenance of gene expression.